3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study not performed to any reported guidance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1968

Report date:

1968

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Not specified

GLP compliance:

no

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol

EC Number:

216-971-0

EC Name:

3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol

Cas Number:

1709-70-2

Molecular formula:

C54H78O3

IUPAC Name:

3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol

Details on test material:

A 100 kg sample of lonox 330, batch number 89/5/6232, purity 98% was supplied by Shell Chemical Company, Carrington.

Test animals

Species:

dog

Strain:

Beagle

Sex:

male/female

Details on test animals or test system and environmental conditions:

26 Beagle hounds, aged about five months and bred at this laboratory were used. The dogs were inoculated at three months with a vaccine consisting of living, modified, tissue culture adapted distemper virus, inactivated canine hepatitis virus and inactivated leptospira bacteria. They were also treated for nematode parasites with piperazine adipate.

Administration / exposure

Route of administration:

oral: feed

Vehicle:

not specified

Details on oral exposure:

Not specified

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not specified

Duration of treatment / exposure:

2 years

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 2000, 10000 ppm
Basis:
nominal in diet

No. of animals per sex per dose:

0 - 5 males & 5 females
2000 - 4 males & 4 females
10000 - 4 males & 4 females

Control animals:

yes, plain diet

Details on study design:

Groups of four males and four females, housed individually, received diets containing 2,000 and 10,000 ppm Ionox 330, an intake of 1 ppm in the dogs used for this experiment being equivalent to 0.05 mg/kg/day.
Control groups, five males and five females, received untreated powdered diet (Powdered 'More-Meat' Dog Food. Quaker Oats Ltd., Southall, Middlesex). Dogs within each of the available litters were allocated at random to the various treatments. Here this allocation was not possible animals of a similar age and weight were used.

Positive control:

Not specified

Examinations

Observations and examinations performed and frequency:

Daily records of general health and behaviour were made and body weights were recorded weekly. Blood samples were taken prior to the experiment. After six and 12 weeks exposure and thereafter at three monthly intervals. Haematological examinations carried out on all blood samples comprised haemoglobin and packed cell volume determinations and erythrocyte, leucocyte and di-ferential leucocyte counts. Clinical chemical determinations were made of serum urea and total protein. Serum protein fractions were determined by paper electrophoresis using LKB paper electrophoresis equipment (LKB buffer pH 8.6, μ = 0.1). Urine examinations and bromsulphthalein (BSP) liver function tests were carried out on the control and top dose groups every four months.

Sacrifice and pathology:

At autopsy, a gross pathological examination was made and the major visceral organs weighed. Histopathological examination was made of haematoxylin and eosin stained sections of liver, thyroid, parathyroid, heart, lungs, spleen, kidneys, adrenals, small and large intestines, stomach, pancreas, skin, skeletal muscle, salivary glands, lymph nodes, bladder, testes, prostate, uterus, ovaries, Fallopian tubes, eye and brain of all animals.

Other examinations:

No data

Statistics:

Not specified

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

Throughout the experiment, the general health, behaviour and body weights of the dogs remained unaffected by the feeding of Ionox 330. During the first 12 weeks of the experiment, one female from each treatment group, all from one litter, became lethargic, anorexic and pyrexial. Other symptoms developed and distemper was diagnosed. These symptoms occurred four weeks after vaccination for distemper, leptospirosis and hepatitis of all puppies in the kennel. As a result of this infection, the 2,000 ppm dog was killed in the fourth week and the 10,000 ppm animal in the 12th week of the experiment. The pathological findings confirmed the clinical diagnosis. During the eighth week, the affected control group female recovered within seven days and showed no further effects.
No changes in organ weights or organ/body weight ratios were observed. The haematology and clinical chemistry of the dogs remained unaffected throughout the exposure, the minor variations observed not being considered of any biological significance.
At autopsy, no gross or microscopic changes attributable to the exposure were found. Lesions associated with ascarid infestation were detected in the livers of both treated and control animals. Evidence of pulmonary and renal disease was also found in all the animals.

Effect levels

Dose descriptor:

NOEL

Effect level:

10 000 ppm

Based on:

test mat.

Sex:

male/female

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1 – Summary of the pathological findings in dogs fed Ionox 330 for two years

1a - Dogs killed during the experiment

Dietary Concn (ppm)	Animal Number	Time of Exposure (weeks)	Gross and microscopic pathological findings
2000	357 Female	4	Focal meningitis, Peri-arteritis of large intestine. Haemorrhage of lymph nodes.
10000	360 Female	12	Bronchopneumonia with numerous abscesses.

1b - After two years

Dietary Concn (ppm)	Animal Number	Sex	Gross and microscopic pathological findings
10000	340	Male	Hepatic granulomas. Nephritis.
	348	Male	Negative.
	351	Male	Hepatic granulomas. Nephritis.
	352	Male	Bronchopneumonia. Pyelonephritis.
	321	Female	Pneumonitis with granulomas. Hepatic granulomas. Nephritis.
	343	Female	Hepatic granulomas.
	356	Female	Hepatic granulomas. Nephritis.
2000	361	Male	Pulmonary fibrosis. Parathyroid adenoma.
	363	Male	Focal pneumonitis.
	339	Male	Hepatic granulomas. Nephritis.
	354	Male	Hepatic granulomas.
	342	Female	Pneumonitis with granulomas. Hepatic ganulomas. Pyelitis.
	338	Female	Nephritis and pyelitis.
	345	Female	Hepatic granulomas. Renal abscess. Resolving pneumonia.
0	347	Male	Hepatic – peripertal inflammation.
	349	Male	Focal pulmonary fibrosis. Fatty infiltration myocardium.
	350	Male	Interstitial pneumonitis with fibrosis. Mild pyelitis.
	353	Male	Bronchitis.
	355	Male	Patchy pneumonitis. Hepatic granulomas.
	359	Female	Hepatic granulomas. Nephritis.
	341	Female	Pneumonitis. Hepatic granulomas. Pyelitis.
	344	Female	Hepatic granulomas. Nephritis.
	346	Female	Patchy pneumonitis. Hepatic granulomas. Nephritis.
	358	Female	Hepatic granulomas.

 

Table 2 – The organ weights and organ/body weight ratios of dogs fed Ionox 330 for two years

Dietary Concn (ppm)	Number of Animals	Terminal body wt (kg)		Organ weight (g)
				Brain	Heart	Liver	Kidneys	Testes
Males
0	5	14.8		83.5	126.1	451	678	16.2
2000	4	15.2		85.4	128.3	487	71	17.2
10000	4	14.9		84.6	118.6	410	67	17.9
S.E. of a treatment mean	5	± 0.61		± 3.28	± 6.76	± 32.5	± 3.2	± 1.05
	4	± 0.69		± 3.67	± 7.55	± 36.3	± 3.6	± 1.05
Females
0	5		12.5	78.3	91.7	349	48
2000	3		13.0	74.8	93.7	332	46
10000	3		13.3	77.3	96.0	349	51
S.E. of a treatment mean	5		± 0.42	± 2.62	± 4.73	± 25.2	± 4.2
	3		± 0.54	± 3.38	± 6.11	± 32.6	± 5.4
				Organ/body weight ration (g/100 g body weight)
Male
0	5			0.57	0.85	3.02	0.45	0.11
2000	4			0.57	0.85	3.21	0.47	0.12
10000	4			0.57	0.80	2.75	0.45	0.12
S.E. of a treatment mean	5			± 0.033	± 0.044	± 0.145	± 0.018	± 0.011
	4			± 0.037	± 0.049	± 0.162	± 0.020	± 0.011
Females
0	5			0.61	0.73	2.77	0.38
2000	3			0.59	0.72	2.54	0.35
10000	3			0.59	0.72	2.63	0.38
S.E. of a treatment mean	5			± 0.013	± 0.028	± 0.135	± 0.021
	3			± 0.016	± 0.036	± 0.174	± 0.027

 

Applicant's summary and conclusion

Conclusions:

In conclusion, this experiment shows Ionox 330 produces no effects when fed to dogs for two years at dietary concentrations of up to 10,000 ppm.

Executive summary:

lonox 330 was fed to dogs for two years at dietary concentrations of 0, 2,000 and 10,000 ppm. Observations were made on the health, behaviour and body weight of all animals. Blood samples were taken regularly for routine haematologica1 and clinical chemical examinations. Liver function tests were carried out at intervals on the control and top dose groups. At autopsy, major visceral organs were weighed and a histopathological examination made of all tissues.

The general health, behaviour and growth rate of the dogs remained unaffected throughout the experiment. No changes were observed in the organ weights and organ/body weight ratios, haematology and clinical chemistry of the animals.No lesion attributable to the exposure to lonox 330 was detected.

A dietary concentration of 10,000 ppm lonox 330 did not produce any toxicological effects when fed to dogs for two years.